Stroke is a leading cause of morbidity and mortality in the United States. However less than 20% of patients are eligible for the current approved interventions of tissue plasminogen activator or thrombectomy. We seek to develop new therapeutics to reduce the extent of damage and functional impairment resulting from ischemic injury to the brain, an area of significant unmet medical need. We have found that interferon regulatory factor (IRF) mediated gene transcription may represent an endogenous mechanism of neuroprotection that is associated with a reduction in ischemic injury. Using both cell and mouse models of stroke we have demonstrated that administration of compounds following the ischemic insult, that are known to induce IRF mediated gene transcription significantly reduces the extent of damage. These results indicate that activation of IRF transcription factors following stroke may be a viable therapeutic intervention for the treatment of stroke patients. The ultimate goal of this STTR program is to identify IRF activators with minimal off-target immune activity and specificity for both mouse and human for preclinical development for stroke. Our specific goal for this Phase I application is to develop and validate a high-throughput screening platform to identify clinically viable IRF activating compounds for further development. We propose the following aims: Aim 1: Validate a high-throughput primary screening assay to identify potent and selective IRF activators using a human THP1 dual ISRE/NF?B reporter cell line and 384-well format. Aim 2: Validate a high-throughput secondary screening assay in mouse J774 cells containing dual ISRE/NFkB reporters to evaluate cross-species activity of primary hits from Aim 1. Aim 3: Verify the target and species selectivity of secondary hits by evaluating a panel of downstream IRF-inducible genes in human and mouse brain-derived cells.